LOCUS Exported 9957 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Express C-terminal ISC mutant POLE1. ACCESSION . VERSION . KEYWORDS pMXS-IRES-Blast POLE1 C-ISCmut SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9957) AUTHORS Sviderskiy VO, Blumenberg L, Gorodetsky E, Karakousi TR, Hirsh N, Alvarez SW, Terzi EM, Kaparos E, Whiten GC, Ssebyala S, Tonzi P, Mir H, Neel BG, Huang TT, Adams S, Ruggles KV, Possemato R TITLE Hyperactive CDK2 Activity in Basal-like Breast Cancer Imposes a Genome Integrity Liability that Can Be Exploited by Targeting DNA Polymerase epsilon. JOURNAL Mol Cell. 2020 Oct 28. pii: S1097-2765(20)30723-1. doi: 10.1016/j.molcel.2020.10.016. PUBMED 33152268 REFERENCE 2 (bases 1 to 9957) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9957 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..417 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 347..369 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 385..401 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 427..801 /label=pol region /note="Moloney murine leukemia virus (MMLV) pol region containing the splice acceptor site" primer_bind 739..755 /label=pBMN 5' /note="MMLV sequence, for inserts in pBMN retroviral vector" CDS 850..915 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" misc_feature 7855..8441 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(8022..8039) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8249..8268 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" LTR 8886..9479 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 9542..9899 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" ORIGIN 1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg 61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca 121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc 181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc 241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta 301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct 361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc 421 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct 481 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta 541 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg 601 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag 661 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc 721 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg 781 ggtggaccat cctctagact gccggatcta gctagttaat taaggatccc taaatcgcaa 841 ttgaccatgg actacaagga ccacgacggt gactacaagg accacgacat cgactacaag 901 gacgacgacg acaagggagg aggcggaggt ggaatgtctc tgaggagcgg cgggcggcgg 961 cgcgcggacc caggcgcgga tggcgaggcc agcagggatg atggcgccac ttcctcagtt 1021 tcggcactca agcgcctgga acggagtcag tggacggata agatggattt gcggtttggt 1081 tttgagcggc tgaaggagcc tggtgagaag acaggctggc tcattaacat gcatcctacc 1141 gagattttag atgaagataa gcgcttaggc agtgcggtag actattattt catccaagat 1201 gacggaagca gatttaaggt ggctttgccc tataaaccgt atttctacat tgcgaccaga 1261 aagggttgtg agcgagaagt ttcatctttt ctctccaaga agtttcaggg caaaattgca 1321 aaagtggaga ctgtccccaa agaggatctg gacttgccaa atcacttggt gggtttgaag 1381 cgaaattaca tcaggctgtc cttccacact gtggaggatc ttgtcaaagt gaggaaggag 1441 atctcccctg ccgtgaagaa gaacagggag caggatcacg ccagcgacgc gtacacagct 1501 ctgctttcca gtgttctgca gaggggcggt gtcattactg atgaagagga aacctctaag 1561 aagatagccg atcaactaga taatatcgtg gacatgcgcg agtacgatgt tccctaccac 1621 atccgcctct ccattgacct gaagatccac gtggctcatt ggtacaatgt cagataccga 1681 ggaaatgctt ttccggtaga aatcacccgc cgagatgacc ttgttgaacg acctgaccct 1741 gtggttttgg catttgacat tgagacgacc aaactgcccc tcaagtttcc tgatgctgag 1801 acagaccaga ttatgatgat ttcctacatg atcgatggcc agggctacct catcaccaac 1861 agggagattg tttcagaaga tattgaagat tttgagttca cccccaagcc agaatatgaa 1921 ggcccctttt gtgtcttcaa tgaacccgat gaggctcatc tgatccaaag gtggtttgaa 1981 cacgtccagg agaccaaacc caccatcatg gtcacctaca acggggactt ttttgactgg 2041 ccatttgtgg aggcccgggc agcagtccac ggtctgagca tgcagcagga gataggcttc 2101 cagaaggaca gccaggggga gtacaaggcg ccccagtgca tccacatgga ctgcctcagg 2161 tgggtgaaga gggacagtta ccttcctgtg ggcagtcata atctcaaggc ggccgccaag 2221 gccaagctag gctatgatcc cgtggagcta gacccggagg acatgtgccg gatggccacg 2281 gagcagcccc agactctggc cacgtattct gtgtcagatg ctgtcgccac ttactacctg 2341 tacatgaagt acgtccaccc attcatcttt gctctgtgca ccattattcc catggagccc 2401 gacgaggtgc tgcggaaggg ctctggcact ctgtgtgagg ccttgctgat ggtgcaggcc 2461 ttccacgcca acatcatctt ccccaacaag caagagcagg agttcaataa gctgacggac 2521 gacggacacg tgctggactc tgagacctac gtcgggggcc acgtggaggc cctcgagtct 2581 ggggttttcc gcagcgatat cccttgccgg tttaggatga atcctgccgc ctttgacttc 2641 ctgctgcagc gggttgagaa gaccttgcgc cacgcccttg aggaagagga gaaagtgcct 2701 gtggagcaag tcaccaactt tgaagaggtg tgtgatgaga ttaagagcaa gcttgcctcc 2761 ctgaaggacg ttcccagccg catcgagtgt ccactcatct accacctgga cgtgggggcc 2821 atgtacccca acatcatcct gaccaaccgc ctgcagccct ctgccatggt ggacgaagcc 2881 acctgtgctg cctgtgactt caataagcct ggagcaaact gccagcggaa gatggcctgg 2941 cagtggaggg gcgagttcat gccagccagt cgcagcgaat accatcggat ccagcaccag 3001 ctggagtcag agaagttccc ccccttgttc ccagaggggc cagctcgggc ctttcatgaa 3061 ctgtcccgcg aggaacaggc gaaatacgag aagagaaggc tggcggatta ctgccggaaa 3121 gcctacaaga agatccacat caccaaggtg gaagagcgtc tcaccaccat ctgccagcgg 3181 gaaaactcct tctacgtgga caccgtgcgt gccttccggg acaggcgtta cgagttcaaa 3241 gggctccaca aggtgtggaa aaagaagctc tcggcggccg tggaggtggg cgacgcggct 3301 gaggtgaagc gctgcaagaa catggaggtg ctgtatgact cgctgcagct ggcccacaag 3361 tgcatcctga actccttcta tggctatgtc atgcgcaagg gggctcgctg gtactccatg 3421 gagatggctg gcatcgtctg cttcacaggg gccaacatca tcacccaggc acgggagctg 3481 atcgagcaga ttgggaggcc cttagagctg gacacagatg gtatatggtg cgtcctgccc 3541 aacagcttcc cagaaaattt tgtcttcaag acgaccaatg tgaagaagcc caaagtgacc 3601 atctcctacc caggcgccat gttgaacatc atggtcaagg aaggcttcac caatgaccag 3661 taccaggagc tggctgagcc gtcctcactc acctacgtca cccgctcaga gaacagcatc 3721 ttttttgagg ttgatgggcc ctaccttgcc atgattcttc cagcctccaa ggaagaaggc 3781 aagaaattga agaagaggta tgctgtgttc aatgaagacg gttctctggc tgagctcaag 3841 ggctttgagg tcaaacgccg cggggaactg cagctgatta agatcttcca atcctcggtg 3901 tttgaggcct tcctcaaggg cagcacgctg gaagaggtgt atggctctgt agccaaggtg 3961 gctgactact ggctggacgt gctgtacagc aaggcagcca acatgcctga ctctgagcta 4021 ttcgagctca tctctgagaa ccgttccatg tctcggaagc tggaagatta cggggagcag 4081 aagtctacgt ccatcagcac agcaaagcgc ctggccgagt tcctgggaga ccagatggtc 4141 aaggatgcag ggctgagttg ccgctacatc atctcccgca agcccgaggg ctcccctgtc 4201 acggagaggg ccatcccact tgccattttc caagcagagc ccacggtgag gaagcacttt 4261 ctccggaaat ggctcaagag ctcttccctt caagactttg atattcgagc aattctggat 4321 tgggactact acattgagcg gctgggaagc gccatccaga agatcatcac catccctgcg 4381 gccctgcagc aggtaaagaa cccagtgcca cgtgtcaaac accccgactg gctgcacaaa 4441 aaactgctgg agaagaatga tgtctacaag cagaagaaga tcagtgagct cttcaccctg 4501 gagggcagga gacaggtcac gatggccgag gcctcagaag acagtccgag gccaagtgct 4561 cctgacatgg aggacttcgg cctcgtaaag ctgcctcacc cagcagcccc tgtcactgtg 4621 aagaggaagc gagttctttg ggagagccag gaggagtccc aggacctcac gccgactgtg 4681 ccctggcagg aaatcttggg gcagcctccc gccctgggaa ccagccagga ggaatggctt 4741 gtctggctcc ggttccacaa gaagaagtgg cagctgcagg cccggcagcg cctcgcccgc 4801 aggaagaggc agcgtctgga gtcggcagag ggtgtgctca ggcccggggc catccgggat 4861 ggtcctgcca cggggctggg gagcttcttg cgaagaactg cccgcagcat cctggacctt 4921 ccgtggcaga ttgtgcagat cagcgagacc agccaggccg gcctgttcag gctgtgggcg 4981 ctcgttggca gtgacttgca ctgcatcagg ctgagcatcc cccgtgtgtt ctacgtgaac 5041 cagcgagtcg ctaaagcgga ggagggtgct tcgtatcgca aggtaaatcg ggtccttcct 5101 cgctccaaca tggtctacaa tctctatgag tattcagtgc cagaggacat gtaccaggaa 5161 cacatcaacg agatcaacgc tgagctgtca gcgccagaca tcgagggcgt atatgagact 5221 caggttccgt tactgttccg ggccctggtg cacctgggct gtgtgtgtgt ggtcaataaa 5281 cagctggtga ggcacctttc aggctgggaa gcagagacct ttgctcttga gcacctggag 5341 atgcgctctc tggcccagtt cagctacctg gaaccaggga gtatccgcca tatctacctg 5401 taccaccacg cacaggccca caaagcgctc ttcgggatct tcatcccctc acagcgcagg 5461 gcatccgtct ttgtgctgga cactgtgcgc agcaaccaga tgcccagcct tggcgccctg 5521 tactcagcag agcacggcct cctcctggag aaggtgggcc ctgagctcct gccacccccc 5581 aaacacacct tcgaagttcg ggcagaaact gacctgaaga ccatctgcag agccatccag 5641 cgattcctgc tcgcctacaa ggaggagcgc cgggggccca cactcatcgc tgttcagtcc 5701 agctgggagc tgaagaggct ggccagtgaa attcctgtct tggaggaatt cccactggtg 5761 cctatctgtg tggctgacaa gatcaactat ggggtcctgg actggcagcg ccatggagcc 5821 cggcgcatga tccgtcacta cctcaacctg gacacctgcc tgtcgcaggc cttcgagatg 5881 agcaggtact ttcacattcc cattgggaac ctaccagagg acatctccac attcggctcc 5941 gacctcttct ttgcccgcca cctccagcgc cacaaccacc tgctctggct gtcccctaca 6001 gcccgccctg acctgggtgg aaaggaggct gatgacaact gtcttgtcat ggagttcgat 6061 gaccaagcca ctgttgagat caacagttca ggctgttact ccacagtgtg tgtggagctg 6121 gaccttcaga acctggccgt caacaccatt ctccagtctc accatgtcaa cgacatggag 6181 ggggccgaca gcatggggat cagcttcgac gtgatccagc aggcctccct ggaggacatg 6241 atcacgggtg gtcaggctgc cagtgccccg gccagctacg atgagacagc cctgtgctct 6301 aacaccttca ggatcctgaa gagcatggtc gtgggctggg tgaaggagat cacccagtac 6361 cacaacatct atgcagacaa ccaggtgatg cacttctacc gctggcttcg gtcgccatcc 6421 tctctgcttc atgaccctgc cctgcaccgc acactccaca acatgatgaa gaagctcttc 6481 ctgcagctca tcgctgagtt caagcgcctg gggtcatcag tcatctacgc caacttcaac 6541 cgcatcatcc tctgtacaaa gaagcgccgt gtggaagatg ccatcgctta cgtggagtac 6601 atcaccagca gcatccattc aaaggagacc ttccattctc tgacaatttc tttctctcga 6661 tgctgggaat ttcttctctg gatggatcca tctaactatg gcggaatcaa aggaaaagtt 6721 tcatctcgta ttcactgtgg actgcaagac tcccagaaag cagggggagc agaggatgag 6781 caggaaaatg aggacgatga ggaggaaaga gatggggagg aggaggaaga ggcggaggaa 6841 tccaacgtgg aggatttact ggaaaacaac tggaacattt tgcagttttt gccacaggca 6901 gcctcctgcc agaactactt cctcatgatt gtttcagcgt acatcgtggc cgtgtaccac 6961 tgcatgaagg acgggctgag gcgcagtgct ccagggagca cccccgtgag gaggaggggg 7021 gccagccagc tctcccagga ggccgagggg gcggtcggag cccttcccgg aatgatcacc 7081 ttctctcagg attatgtcgc aaatgagctc actcagagct tcttcaccat cactcagaag 7141 attcagaaga aagtcacagg ctctcggaac tccactgagc tctcagagat gtttcctgtc 7201 ctccccggtt cccacttgct gctcaataac cctgccctgg agttcatcaa atacgtgtgc 7261 aaggtgctgt ccctggacac caacatcaca aaccaggtga ataagctgaa ccgagacctg 7321 cttcgcctgg tggatgtcgg cgagttctcc gaggaggccc agttccgaga cccctgccgc 7381 tcctacgtgc ttcctgaggt catctgccgc agctgtaact tctgccgcga cctggacctg 7441 tgtaaagact cttccttctc agaggatggg gcggtcctgc ctcagtggct ctgctccaac 7501 tgtcaggcgc cctacgactc ctctgccatc gagatgacgc tggtggaagt tctacagaag 7561 aagctgatgg ccttcaccct gcaggacctg gtcagcctga agagccgcgg ggtgaaggag 7621 accagcatgc ctgtgtacag cagcagcgcg ggagacttcg ccctcaccat ccacacccag 7681 gtcttcatgg aacagatcgg aatattccgg aacattgccc agcactacgg catgtcgtac 7741 ctcctggaga ccctggagtg gctgctgcag aagaacccac agctgggcca ttaggtcgag 7801 ctaaatcggt ttaaacgcgc taatgcggcc gctacgtaaa ttccgccccc cccccccctc 7861 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt 7921 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc 7981 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca 8041 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac 8101 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg 8161 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt 8221 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct 8281 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg 8341 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt 8401 ggttttcctt tgaaaaacac gatgataata tggccacaac catggttcct ttgtctcaag 8461 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag 8521 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg 8581 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg 8641 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga 8701 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag 8761 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt 8821 atgtgtggga gggctaagtc gacgataaaa taaaagattt tatttagtct ccagaaaaag 8881 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc 8941 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat 9001 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 9061 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 9121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 9181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 9241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 9301 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc 9361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 9421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 9481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 9541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 9601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 9661 ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc 9721 gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc 9781 ccctaatagg agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct 9841 ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc 9901 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattag //